User:Anthony Salvagno/Notebook/Research/2009/08/11/Getting Definitive Unzipping

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Koch wants me to get Definitive unzipping and so do I. Remember I made a challenge to myself to have it done by next Fri. Well he gave me some small tasks that will help me realize it (hopefully).

Current Tethers

To try and debug our current tethers, there are a couple things I will work on today and tomorrow:

  • make new BGB - it is possible our BGB has gone bad and as such I will need to make more.
  • use kappa casein instead of BGB
  • use higher concentrations of DNA

New Tethers

I will make brand new tethers. This is more relatively long term because I need some supplies. In short I will use pBluescript, digest it with SapI or EarI, and ligate it to the unzipping construct. Then I will cap it with a Sap/EarI hairpin to prevent concatamers from forming.

Fork You

I will also try and get the fork DNA to unzip. This is new to my notebook and I will dedicate another post to it, but I need to get this working so we can send some to Doug Smith. This could have to do with our BGB problem.